Biochemical detection of capsid in the nucleus during HIV-1 infection.
Biochemical detection of capsid in the nucleus during HIV-1 infection.
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DOI：
10.1016/j.xpro.2021.100323
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发表时间：
2021-03-19
期刊：
影响因子：
--
通讯作者：
Diaz-Griffero F
中科院分区：
文献类型：
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作者：
Selyutina A;Diaz-Griffero F
为了了解HIV-1衣壳在病毒复制中的作用，我们开发了一种在感染过程中在细胞核中生化追踪衣壳的方案。为此，我们将HIV-1感染的细胞分离成核和胞质组分。通过蛋白质印迹法分析级分的HIV-1衣壳含量以及核和胞质标记物，以评估级分的真实来源。该方案可以应用于循环和非循环人类细胞。有关本方案使用和执行的完整详细信息，请参阅。在人类细胞系中感染期间追踪细胞核中HIV-1衣壳的方法HIV-1感染的细胞核和胞质组分的分析小分子PF 74阻止HIV-1衣壳进入细胞核这种方法可用于循环和非循环人类细胞为了了解HIV-1衣壳在病毒复制中的作用，我们开发了一种在感染过程中生化追踪细胞核中衣壳的方案。为此，我们将HIV-1感染的细胞分离成核和胞质组分。通过蛋白质印迹法分析级分的HIV-1衣壳含量以及核和胞质标记物，以评估级分的真实来源。该方案可以应用于循环和非循环人类细胞。
To understand the role of the HIV-1 capsid in viral replication, we developed a protocol to biochemically track capsid in the nucleus during infection. To this end, we separated HIV-1-infected cells into nuclear and cytosolic fractions. Fractions were analyzed by western blotting for HIV-1 capsid content as well as for nuclear and cytosolic markers to assess the bona fide origin of the fractions. This protocol can be applied in both cycling and non-cycling human cells. For complete details on the use and execution of this protocol, please refer to. Method to track the HIV-1 capsid in the nucleus during infection in human cell lines Analysis of HIV-1 infected nuclear and cytosolic fractions The small molecule PF74 prevents entry of HIV-1 capsid into the nucleus This method can be used in cycling and non-cycling human cells To understand the role of the HIV-1 capsid in viral replication, we developed a protocol to biochemically track capsid in the nucleus during infection. To this end, we separated HIV-1-infected cells into nuclear and cytosolic fractions. Fractions were analyzed by western blotting for HIV-1 capsid content as well as for nuclear and cytosolic markers to assess the bona fide origin of the fractions. This protocol can be applied in both cycling and non-cycling human cells.